Determination of urinary thymidine glycol using affinity chromatography, HPLC and post-column reaction detection : a biomarker of oxidative DNA damage upon kidney transplantation

Thier, Ricarda, Brüning, Thomas, Kocher, Kristin, Blaszkewicz, Meinolf, Makropoulos, Vassilios, Sundberg, Anders, & Bolt, Hermann M. (1999) Determination of urinary thymidine glycol using affinity chromatography, HPLC and post-column reaction detection : a biomarker of oxidative DNA damage upon kidney transplantation. Archives of Toxicology, 73(8-9), pp. 479-484.

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Abstract

Reactive oxygen species are generated during ischaemia-reperfusion of tissue. Oxidation of thymidine by hydroxyl radicals (HO) leads to the formation of 5,6-dihydroxy-5,6-dihydrothymidine (thymidine glycol). Thymidine glycol is excreted in urine and can be used as biomarker of oxidative DNA damage. Time dependent changes in urinary excretion rates of thymidine glycol were determined in six patients after kidney transplantation and in six healthy controls. A new analytical method was developed involving affinity chromatography and subsequent reverse-phase high-performance liquid chromatography (RP-HPLC) with a post-column chemical reaction detector and endpoint fluorescence detection. The detection limit of this fluorimetric assay was 1.6 ng thymidine glycol per ml urine, which corresponds to about half of the physiological excretion level in healthy control persons. After kidney transplantation the urinary excretion rate of thymidine glycol increased gradually reaching a maximum around 48 h. The excretion rate remained elevated until the end of the observation period of 10 days. Severe proteinuria with an excretion rate of up to 7.2 g of total protein per mmol creatinine was also observed immediately after transplantation and declined within the first 24 h of allograft function (0.35 + 0.26 g/mmol creatinine). The protein excretion pattern, based on separation of urinary proteins on sodium dodecyl sulphate-polyacrylamide gel electrophorosis (SDS-PAGE), as well as excretion of individual biomarker proteins, indicated nonselective glomerular and tubular damage. The increased excretion of thymidine glycol after kidney transplantation may be explained by ischaemia-reperfusion induced oxidative DNA damage of the transplanted kidney.

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6 citations in Web of Science®

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ID Code: 77480
Item Type: Journal Article
Refereed: Yes
Keywords: DNA damage, Kidney transplantation, Reactive oxygen species, Renal ischaemia, Thymidine glycol, 5, 6 dihydro 5, 6 dihydroxythymidine, cell DNA, reactive oxygen metabolite, adult, affinity chromatography, analytic method, article, clinical article, clinical trial, controlled clinical trial, controlled study, diagnostic accuracy, diagnostic value, high performance liquid chromatography, human, kidney allograft, kidney allograft rejection, male, priority journal, proteinuria, reperfusion injury, urinalysis, urinary excretion, validation process, Biological Markers, Chromatography, Affinity, Chromatography, High Pressure Liquid, Electrophoresis, Polyacrylamide Gel, Female, Humans, Kidney Glomerulus, Kidney Tubules, Middle Aged, Thymidine
DOI: 10.1007/s002040050638
ISSN: 0340-5761
Divisions: Current > Schools > School of Clinical Sciences
Current > QUT Faculties and Divisions > Faculty of Health
Copyright Owner: Copyright 1999 Springer-Verlag
Deposited On: 16 Oct 2014 04:35
Last Modified: 16 Oct 2014 04:44

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